1. Field Of The Invention
The present invention is related to a novel hemostatic composition. More specifically, the present invention is related to a hemostatic composition comprising thrombin, casein kinase II, and sphingosine or a sphingosine derivative and providing rapid clotting and hemostasis.
2. Description Of The Prior Arts
Hemostasis involves three complex mechanisms: clot formation, rapid constriction of the injured blood vessel and the aggregation of platelets to form a plug on the injured surface of the blood vessel. A clot is formed by a series of transformations involving more than ten different proteins, calcium ion (Ca2+) and thromboplastin. When bleeding occurs, fibrinogen, which is highly soluble, is converted into insoluble fibrin monomer by the proteolytic action of thrombin. The fibrin monomers spontaneously associate to form a clot, on which factor XIIIa acts to aggregate platelets to form a plug on the injured surface of the blood vessel.
It has been reported that phosphate groups attached to fibrinogen affect the gelation of thrombin (Forsberg, P. O., Thromb. Res., 53, 1-9, 1989). And, various protein kineses such as protein kinase A (Engstrim, L., Edlund B., Rangnarsson, U., Dahlqvist-Edberg, U., and Humble, E., Biochem. Biophys. Res. Commun., 96, 1507, 1980), protein kinase C, casein kinase I (Itarte, E., Plana, M., Guasch, M. D., and Martos, C., Biochem. Biophys, Res. Commun., 117, 631-636, 1983), and casein kinase II (Humble, E., Heldin, P., Forsberg, P. O. and Engstrom, L., Arch, Biochem. Biophys., 241, 225-231, 1985) have been reported to be involved in the phosphorylation of fibrinogen. The effect of the phosphorylation of fibrinogen on the thrombin-induced gelation of fibrinogen varies depending on thekinase used to phosphorylate fibrinogen. The phosphorylated fibrinogen is ineffeciently cleaved by plasmin regardless of the kind of the kinase involved in the phosphorylation thereof (Martin, S. C., et al., Thromb. Res. 61, 243252, 1991).
Sphingosine, which is one kind of sphingolipid, has important roles in the growth and differentiation of cells. Further, sphingosine also is reported to have an ability to increase the activity of casein kinase II (McDonald, O.Bradley; Hannun, Yusuf A.; Reynolds, E. Hugh and Sahyoun, Naji, Journal of Biological Chem., Vol. 266, No. 32, pp 21773-21776, 1991).
A property required to serve as an efficient hemostatic agent is fast control of bleeding in order to prevent entry of pathogens at the bleeding site or massive hemorrhage during operations. Thrombin preparations have been most widely employed as commercially available hemostatic compositions.
The inventors conducted extensive studies to develop a more efficient hemostatic composition, which allows a faster hemostasis than conventional hemostatic compositions. As a result thereof, they surprisingly found that, if the thrombin-containing hemostatic compositions contains casein kinase II, a protein phosphorylation enzyme, together with sphingosine or a sphingosine derivative, it can exhibit its hemostatic activity in a shorter period of time.